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Contrary to prior speculations, a fluorescent scan unambiguously shows that TphB coordinates Zn2 + and not Fe2 +.
The concentrated protein is then loaded onto a size-exclusion chromatography column to remove the remainder of the free dye. Figure 2 shows the removal of the free dye from the sample as well as the separation of multimer forms of the protein from monomer as visualized with a fluorescent scan of the SDS-PAGE using a Typhoon imager.
(A) Fluorescent scan of Hoechst-stained cells in VitroTube (transmitted light overlay).
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The array sites containing probe DNA and the control sites without probes are readily distinguishable using laser-assisted fluorescent scanning.
The GDNF was then allowed to diffuse out of the scaffold, and release was tracked via fluorescent scanning confocal microscopy.
A simple method for separation and quantification of neutral lipids was developed using thin-layer chromatography (TLC) and high-performance fluorescent scanning.
Data were quantified using Microarray Imager or ElectraSense software (CombiMatrix) for fluorescent scans or ECD respectively.
Treated samples were then washed 5 times with PBS and immediately visualized by live confocal fluorescent scanning microscopy.
The processed microarrays were visualized by fluorescent scanning and the resulting images were quantified as described in [13].
To allow similar analysis of unopsonized phagocytosis of specific bacterial pathogens we have developed high-throughput fluorescent scanning cytometry-based methods for quantification of phagocytosis of S. aureus by adherent human alveolar-like macrophages in a 96-well microplate format.
The protein pellet was subsequently resuspended in 2X SDS-protein reducing buffer and boiled for 10 min. Proteins were then separated by SDS-PAGE analyzedyzed by in-gel fluorescent scanning using a Typhoon scanner (GE Healthcare; excitation at 532 nM, emission at 580 nM).
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